Methodology for the isolation and purification of vesicles, i.e. periplasmic bodies from protoplasted Saccharomyces rouxii will be improved and extended. More uniform starting cells will be sought by centrifugation methods and by induction of synchronous growth in cultures. The latter approach will also be used to examine the periods in the cell cycle in which periplasmic bodies are generated, and in which prospective marker enzymes (acid phosphatase and beta-fructofuranosidase) are synthesized. Purified periplasmic bodies will be analyzed for marked enzyme content as compared with whole cells; and for other organelles will also be assayed and compared. The ultrastructural localization of acid phosphatase by the Gomori reaction will be further refined in its application to yeast cells. The information and methodology obtained with S. rouxii will be applied and extended to studies with Candida albicans and Histoplasma capsulatum. Pretreatments with sub-lethal doses of anti-mycotics will be included. Attendant periplasmic bodies will be examined for possible enrichment in the concentration of polyene complexed with sterol. BIBLIOGRAPHIC REFERENCES: Arnold, W.N., and J.S. Lacy. 1977. Plasmolysis in yeast. Abstract I-125, 77th Annual Meeting of the American Society for Microbiology, New Orleans, Louisiana. Arnold, W.N., and J.S. Lacy. 1977. Permeability of the cell envelope and osmotic behavior in yeast. J. Bacteriol. August, in press.